Advertisement
/ 24 []

1- Making Density Gradients - PowerPoint Slideshow


Discontinuous gradient by overlayering. Discontinuous gradient by underlayering. . Diffusion of a discontinuous gradient. Rapid formation of continuous from discontinuous gradient. Two-chamber device for continuous gradients. The Gradient Master. B. . Gradient Master profiles from 10% and 40% iodixanol at 80

Download Presentation

1- Making Density Gradients

An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use only and may not be sold or licensed nor shared on other sites. SlideServe reserves the right to change this policy at anytime.While downloading, If for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.











- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -




Presentation Transcript


1 making density gradients l.jpg

1- Making Density Gradients

  • Pre-formed discontinuous gradients

  • Pre-formed continuous gradients

  • Self-generated gradients

2- Density Gradient Harvesting


Discontinuous gradient by overlayering l.jpg

C

A

B

Discontinuous gradient by overlayering


Discontinuous gradient by underlayering l.jpg

A

B

C

D

Discontinuous gradient by underlayering


Diffusion of a discontinuous gradient l.jpg

density

top

bottom

Diffusion of a discontinuous gradient


Rapid formation of continuous from discontinuous gradient l.jpg

Discontinuous

Continuous

45-60 min

Rapid formation of continuous from discontinuous gradient


Two chamber device for continuous gradients l.jpg

A

B

T

SB

SB

P

M

Two-chamber device for continuous gradients


The gradient master l.jpg

B

The Gradient Master


Gradient master profiles from 10 and 40 iodixanol at 80 and 20 rpm effect of time l.jpg

Density (g/ml)

2 min

1.13

2.5 min

3.0 min

1.11

1.09

1.07

1

3

5

7

9

Fraction Number

Gradient Master profiles from 10% and 40% iodixanol at 80° and 20 rpm: effect of time


Swinging bucket rotor l.jpg

axis of rotation

g

At rest

Spinning

g

Swinging-bucket rotor


Fixed angle and vertical rotors l.jpg

Fixed-angle

Vertical

g

1g

Fixed-angle and vertical rotors


Sedimentation path length of rotors l.jpg

Swinging-bucket

Axis of

rotation

Vertical

Sedimentation path length of rotors


Sample gradient in vertical rotor l.jpg

A

B

Axis of rotation

Sample/gradient in vertical rotor


Self generated gradients l.jpg

time

g

Self-generated gradients

  • At 360,000gav iodixanol molecules will start to sediment


Self generated gradient strategy l.jpg

350,000g

350,000g

1g

1g

Self-generated gradient strategy


Near vertical rotors are best of all l.jpg

Near-vertical rotors are best of all


Iodixanol self generated gradient requirements l.jpg

Iodixanol self-generated gradient requirements

  • Rotors with a short sedimentation path length ~ 17 mm

  • RCF of 180,000-350,000gav


Effect of time on iodixanol self generated gradient in the beckman tln100 at 365 000g l.jpg

Density (g/ml)

15% iodixanol

1.30

1 h

3 h

1.25

20% iodixanol

1.20

1 h

3 h

1.15

1.10

1.05

1.00

1

3

5

7

9

11

13

15

Fraction Number

Effect of time on iodixanol self-generated gradient in the Beckman TLN100 at 365,000g


Effect of rcf and iodixanol in beckman vti65 1 3 h l.jpg

1.3

15% (170,000g)

30% (170,000g)

15% (353,000g)

1.2

30% (353,000g)

Density (g/ml)

1.1

1.0

1

3

5

7

9

11

Fraction Number

Effect of RCF and [iodixanol] in Beckman VTi65.1 (3 h)


Removal of banded material using a syringe l.jpg

Removal of banded material using a syringe


Collecting a band from a sealed tube l.jpg

Collecting a band from a sealed tube


Harvest gradient into equal volume fractions by tube puncture l.jpg

Gilson Fraction

Collector

P

96-well plate

Harvest gradient into equal volume fractions by tube puncture


Upward displacement l.jpg

Perfluorodecalin

Perfluorodecalin

P

P

Upward displacement


Labconco auto densi flow collects from meniscus l.jpg

P

P

P

P

M

B

EP

C

1

2

3

4

Labconco Auto Densi-flow collects from meniscus


Labconco auto densi flow linked to a gilson fc205 fraction collector l.jpg

MB

Labconco Auto Densi-flow linked to a Gilson FC205 Fraction Collector